Objective To study the effect of the products of entermorpha (EP), chlorella growth factor (CGF) and entermorpha dietary fiber (EPDF) on defecation and blood lipid regulation. Method Based on the composition of EP products, the male ICR mice were intervened with EP, EPDF and CGF with three doses. Using the constipation model induced by compound diphenoxylate, the effect of EP products on enterocinesia function of mice was determined by measuring defecation and the percentage of ink-pushing in small intestine. High-lipid rat model was established by feading high fat diet and the intervention effect with three doses of EPDF and CGF for 6 w was observed. Results As compared to the model group,all doses of EP,both middle and high doses of EPDF,and high dose of CGF could shorten the time of first defecation and increase the percentage of ink-pushing in small intestine of costive mice (P

ObjectiveTo study the expression of EZH2 gene in cervical squamous carcinoma and its clinicopathologic significance. MethodsThe expression of EZH2 mRNA was detected in 21 samples of normal cervical tissue, 27 samples of CIN tissue and 48 samples of cervical squamous carcinoma tissue by RT-PCR. And the relationship between EZH2 expression and the clinical pathological characteristic was analyzed. ResultsThe level of EZH2 mRNA in cervical squamous carcinoma tissues(1.67±0.01)were significantly higher than that in the normal cervical tissues (0) and CIN tissues (0.36±0.02) (P ＜ 0.01).There was no correlation between EZH2 and ages (P ＞ 0.05), while the expression of EZH2 was highly correlated with histologic stage,clinical stage,muscular invasion depth and lymph node metastasis of cervical squamous carcinoma (P ＜ 0.01).ConclusionThe over-expression of the EZH2mRNA may play an important role in the pathogenesis and progression of cervical squamous carcinoma,suggesting that EZH2 might be a new biomarker for diagnosing cervical squamous carcinoma.

Objective To detect the expression of heparanase (HPA) in cervical cancer cells and investigate the impact of quercetin on the expression of HPA,and the molecular mechanism that quercetin inhibits the growth of cervical cancer cells.Methods The experimental groups included cervical cancer cell lines (HeLa and Caski) exposed to different concentrations of quercetin (20,40 and 80 μmol/L) in the culture medium.The control groups included a negative control group,which was cultured with RPMI 1640 only,and a positive control group,in which cervical cancer cells were transfected with HPA small interference RNA (siRNA) to silence HPA expression.The cellular expression levels of HPA were detected with fluorescence quantitative real-time PCR and western blot analysis at 24,48 and 72 hours after treatment.Results (1) HPA was significantly expressed in both cervical cancer cell lines (HeLa and Caski),and it exists both nucleus and cytoplasm.(2)The real-time PCR shows as follows:as the quercetin concentration increased (20,40 and 80 μmol/L),the mRNA expression level of HPA decreased (P ＜0.01),in which the inhibition of HPA expression was concentration dependent.In addition,the inhibition of HPA expression was also time dependent.As time growth,the expression level of HPA mRNA (24,48 and 72 hours) in HeLa and Caski cells decreased (P ＜ 0.01).Compared with negative control group,the expression level of HPA mRNA decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P ＜ 0.05) ; Compared with positive control group,the expression level of HPA mRNA expressed no obvious difference in quercetin (80 μmol/L) group (P ＞ 0.05) in HeLa cells,while it was opposite in Caski cells(P ＜0.01).(3)The result of western blot shown that,as the quercetin concentration increased(20,40 and 80 μmol/L)and time growth (24,48 and 72 hours),the expression level of HPA protein decreased (P ＜ 0.01),and the inhibition of HPA protein expression was concentration and time dependent.Compared with negative control group,the expression level of HPA protein decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P ＜ 0.05) ;Conpared with positive control group,the expression level of HPA protein expressed no obvious difference in quercetin (80 μmol/L) group (all P ＞ 0.05) in both HeLa cells and Caski cells (all P＞0.05).Conclusion Quercetin could inhibits the expression of HPA in cervical carcinoma cell lines,which inhibition is concentration and time dependent.

Objective Design and synthesize short hairpin RNA (shRNA) expression vector of RNA for specific silencing of heparanase (HPA) gene,screened plasmid which silence effects is the best.Observe the function of ceil invasion after inhibiting the expression of HPA in cervical carcinoma cell lines (HeLa).Methods The genomic sequence of HPA gene was retrieved from GenBank database.Designed four pairs of specific oligonucleotide sequences and a negative control according to the shRNA design principles.They were inserted into the vector pYr-1.1,vectors,and transfected into HeLa cells via lipofectamine.Reverse transcription (RT)-PCR and immunofluorescence were employed to detect the expression of HPA gene in the transfected cells at the mRNA and protein levels,respectively.The plasmid were screened and transfected into HeLa cells,then transwell small room stromal invasion experiment were employed to observe the cervical carcinoma cell invasion.Results RT-PCR results of transfected HeLa cells shown that the mRNA amplification multiples were 0.54 ±0.05 in the HPA-592 group,0.89 ±0.18 in HPA-995 group,0.82 ±0.22 in the HPA-1351 group,0.91 ±0.47 in HPA-1658 group.While,they were 1.31 ±0.72 and 1.09 ±0.16 in negative control and blank control group,respectively.Green fluorescence was visible in the cytoplasm,which indicated that the HPA protein was expressed in the cytoplasm,of them the weakest green fluorescence in the HPA-592 group.The relative numbers of invasive cells among the HeLa cells were as follows:182 ±6 in the blank control group,258 ± 17 in the negative control group,and 44 ± 4 in the HPA-592-specific interference group(P ＜ 0.01).Conclusion Successfully screened shRNA vector targeting human HPA,efficiently inhibit expression of HPA gene when transfected into HeLa cells,and significantly reduced the invasion capacity of cervical carcinoma cells.

Objective To study the efficacy and safety of Gonadotropin-releasing hormone agonists (GnRH-a) combined with laparoscope conservative surgery in treatment of moderate or severe endometriosis.Methods From Jan.2007 to Jan.2010,68 patients with moderate or severe undergoing treatment in Renmin Hospital of Wuhan University were enrolled in this retrospective study.Three groups were classified,which were 25 patients in GnRH-a group,subcutaneous injection Leuprorelin on the second day of menstruation,every 4 weeks for 3 months.Twenty-three patients in Marvelon group,orally one marvelon tablet on the second day of menstruation,continuous 21 days for one period of treatment for 3 courses.Twenty patients in surgery group,without any medicine used preoperatively.All patients were followed by 12 months and compare their surgery time,blood loss,recovery,visual analog scale (VAS),and recurrence and so on.Results The operating time were (68 ± 18) min in GnRH-a group,(80 ± 21) min in Marvelon group and (90± 24) min in surgery group.The amount of bleeding were (118 ± 15) ml in GnRh-a group,(161 ± 18) ml in Marvelon group and (193 ± 13) ml in surgery group.There was significant lower in the operating time and amount of bleeding in GnRH-a group than those in other two groups (P ＜ 0.05).The activity time and the anus exhaust time were shorter in patients in GnRh-a group than those in the other two groups significantly(P ＜ 0.05).When followed up in 12 months after treatment,visual analogue scale had dropped from 3.8 (1.9-6.8) to 1.9 (1.1-2.8) in GnRh-a group,from 2.7 (1.3-5.5) to 1.8 (1.2-3.2) in Marvelon group and from 1.9(1.0-4.9) to 1.6(1.0-3.6) in surgery group.It was showed the most remarkable decreased VAS in GnRHa group when compared with the other two groups(P ＜ 0.05).The recurrence rates were 12％ (3/25) in GnRH-a group,22％ (5/23) in Marvelon group and 25 ％ (5/25) in surgery group.It was found that the most significant lower recurrence was in GnRH-a group when compared with the other two groups (P ＜ 0.05).Conclusions It was safe and efficacy that GnRH-a combined with laparoscopic conservative surgery were used in treatment of endometriosis.It could bring shorter operation time,less intraoperative blood loss,quick postoperative recover,the lower recurrence rate.

Objective To evaluate the clinical value of adriamycin injection via the foramen ovale and around peripheral trigeminal branches under the guidance of X-ray for treatment of primary trigeminal neuralgia by comparison with the three-dimensional computed tomography (CT).Methods A total of 91 patients with primary trigeminal neuralgia of both sexes,aged 33-76 yr,with the course of disease 6 months-24 yr,with visual analogue scale score of 6-9,were divided into 2 groups using a random number table:X-ray group (n =43) and CT group (n =48).Hartel anterior approach was used to puncture the foramen ovale in 2 groups.One point five percent adriamycin 0.2,0.3 and 0.5 ml were injected via the supraorbital foramen,infraorbital foramen and oval foramen.When pain relief was poor (visual analogue scalc scorc≥ 4) within 1 yr after treatment,oxcarbazepine and adjuncts (tramadol,flupentixol and melitracen tablets,etc.) were taken orally.The requirement for oxcarbazepine and adjuncts was recorded during 1 day-1 week,1 week-1 month,1-3 months,3-6 months and 6 months-1 yr after treatment periods.The operation time,the nuinber of puncture,and developinent and recurrence of complications during treatment and within 1 yr after treatment were recorded.Results Compared with CT group,the number of puncture and incidence of facial hematoma during treatment were significantly increased (P ＜ 0.05 or 0.01),and no significant change was found in the operation time,requirement for oxcarbazepine and adjuncts,incidence of dizziness,nausea and vomiting during treatment,or the incidence and recurrence rate of masticatory muscle weakness and facial numbness after treatment in X-ray group (P＞0.05).Conclusion Compared with the three dimensional CT,X-ray provides similar efficacy and safety when used to guide adriamycin injection via the foramen ovale and around peripheral trigeminal branches for treatment of primary trigeminal neuralgia,showing that X-ray guidance has significant clinical value.

Objective To find out the association between the indicators(pulse concussion lung function test index) of bronchial hyperresponsiveness (BHR) with fractional concentration of exhaled nitric oxide (FeNO) at different control periods among preschool asthmatic children.Methods Totally 74 asthmatic children in the pediatric department of our hospital from April 2015 to February 2017 were enrolled in this study,and 25 children undergoing the lung function and FeNO examination served as the controls,aged 3-5 years old.The cases were divided into three groups according to the standard in 2016 version of the Prevention and Treament Guide of Children Bronchial Asthma:asthma control group(n =26),asthma non-control;group(n =48) and control group (n=25).All data of FeNO,resistance of the respiratory system at 5 Hz(R5),resistance of the respiratory system at 5 Hz (R20),difference of R5 and R20(R5-20),reactance area (AX),reactance of the respiratory system at 5 Hz (X5) and resonant frequency of reactance (Fres) were collected.The FeNO,pulse concussion lung function test value and their association were analyzed.Results (1) The FeNO value of asthma the non-control group was significantly higher than that of the asthma control group and the control group,which were 34.00 ± 18.17,20.23± 11.07 and 28.00± 17.30 respectively.The AX detection value of the asthma non-control group was significantly higher than that of the control group(37.29 ± 15.27 vs.30.17 ± 9.50,P＜0.05).(2)R20 had weak correlation with FeNO in the control group(P＜0.05),while R20 had no correlation with FeNO in the non-control group and control group (P＞0.05).FeNO had no obvious correlation with R5,R520,AX,X5 and Fres in the asthma non-control group,asthma control group and control group(P＞0.05).Conclusion In preschool children with asthma,FeNO can reflect the airway eosinophilic inflammation control,and does not reflect the airway hyperresponsiveness.Thereforeit ie needed to combined with FeNO and IOS indicators (airway hyperresponsiveness index AX,etc.),which can more precisely judge whether asthma being controlled.

Objective To compare expression of Toll?like receptors 7 and 9(TLR7, TLR9)as well as their regulatory molecules myeloid differentiation factor 88(MyD88)and nuclear factor?κB(NF?κB)in peripheral blood mononuclear cells(PBMCs)between patients with vitiligo and healthy individuals, and to explore their significance. Methods Flow cytometry was performed to measure expression of TLR7 and TLR9 in PBMCs among 36 patients with vitiligo and 22 healthy controls, and real?time fluorescence?based quantitative PCR(RT?PCR)was conducted to determine mRNA expression of MyD88 and NF?κB in the above blood samples. Results Compared with healthy controls, patients with vitiligo showed higher expression of TLR7 and mRNA expression of MyD88 and NF?κB, but lower expression of TLR9. However, significant differences were only observed in the mRNA expression of NF?κB(t=2.814, P=0.008), but not in the expression of TLR7 and TLR9 or the mRNA expression of MyD88 between patients and controls (t = 1.477, 1.761, 0.058, all P > 0.05). Conclusion NF?κB, as a key signaling molecule of TLR7 and TLR9 regulation pathways, increases obviously in patients with vitiligo, suggesting that NF?κB may be involved in the pathogenesis of vitiligo.

A cDNA, named Dd-ace-2, encoding an acetylcholinesterase (AChE, EC3.1.1.7), was isolated from sweet-potato-stem nematode, Ditylenchus destructor. The nucleotide and amino acid sequences among different nematode species were compared and analyzed with DNAMAN5.0, MEGA3.0 softwares. The results showed that the complete nucleotide sequence of Dd-ace-2 gene of Ditylenchus destructor contains 2425 base pairs from which deduced 734 amino acids (GenBank accession No. EF583058). The homology rates of amino acid sequences of Dd-ace-2 gene between Ditylenchus destructor and Meloidogyne incognita, Caenorhabditis elegans, Dictyocaulus viviparous were 48.0%, 42.7%, 42.1% respectively. The mature acetylcholinesterase sequences of Ditylenchus destructor may encode by the first 701 residues of deduced 734 amino acids.The conserved motifs involved in the catalytic triad, the choline binding site and 10 aromatic residues lining the catalytic gorge were present in the Dd-ace-2 deduced protein. Phylogenetic analysis based on AChEs of other nematodes and species showed that the deduced AChE formed the same cluster with ACE-2s.
Acetylcholinesterase ; genetics ; Amino Acid Sequence ; Animals ; Cloning, Molecular ; DNA, Complementary ; genetics ; Genes, Helminth ; genetics ; Ipomoea batatas ; parasitology ; Molecular Sequence Data ; Nematoda ; enzymology ; genetics ; Plant Stems ; parasitology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, Protein

Objective To prepare and characterize salinomycin sodium‐loaded nano liposomes(SLN) .Methods The nano liposomes were prepared by a thin‐film dispersion method .The formula of SLN was optimized by regulating the cholesterol ratio of the nano liposomes ,using the encapsulation efficacy (EE) of SLN as the primary outcome measure .Results Transmission e‐lectron microscope (TEM) showed that SLN was round and had a good dispersion .Dynamic laser scatter (DLS) showed that SLN was of a desired size of 99 nm ,and zeta potential of -33 .5 mV .EE of SLN was 85 .7% and drug loading of 6 .7% .Ac‐cording to the formulation of nano liposomes ,the concentration of salinomycin sodium in water was greatly improved by 15 folds .Additionally ,the nano liposomes were observed to exhibit sustained release characteristics .Conclusion Salinomycin sodi‐um‐loaded nanoliposomes of a desired size of about 100 nm were obtained ,which were well dispersion ,and high EE and drug loading .Solid pharmaceutics foundation for the activity examination of SLN was provided in this research .